Histopathological alterations on rat after exposed to deltamethrin by inhalation treatements

THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY NGUYEN THI VAN HISTOPATHOLOGICAL ALTERATIONS ON RAT AFTER EXPOSED TO DELTAMETHRIN BY INHALATION TREATEMENTS BACHELOR THESIS Study Mode: Full-time Major : Environmental Science and Management Faculty : International Training and Development Center Batch : 2012-2016 Thai Nguyen, 05/12/2016 DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Degree Program : Bachelor of Environmental Science and Management Student name : Nguyen Thi Van Student ID : DTN1253150042 Thesis Title : HISTOPATHOLOGICAL ALTERATIONS ON RAT AFTER EXPOSED TO DELTAMETHRIN BY INHALATION TREATEMENTS Supervisor (s): Dr Nguyen The Hung Dr Arinafril Dr Krisna Dr.Mulawarman Abstract: Deltamethrin is a synthetic pyrethroid widely used as the insecticide of choice especially for local vector mosquitoes in most countries This experimental study aimed to evaluate the morphologic changes in the lungs caused by the inhalation of this insecticide The study was performed on nine Sprague-Dawley rats There were three rats which were used as control group (without Deltamethrin inhalation), the other six rats were used as treated group which was exposed to 1:10 dilution of deltamethrin aerosol spray for hour each day After 8, 16, 24 days, a group of three rats (including control and two-treated rats) was sacrificed and tissue samples taken from the lungs were processed for both light microscopy and transmission electron ii microscopy This experiment was finished after 21 days Light microscopic examination revealed heavy congestion, marked perivascular edema, and lymphoplasmocytic infiltration with focal nonspecific interstitial pneumonia, foamy macrophage accumulation, emphysema, peribronchial lymphoid tissue hyperplasia, and focal hemorrhage Ultrastructurally, the ciliated cells of the airways appeared swollen with a few structurally abnormal cilia Alveolar lining cells revealed mild degeneration and a slight hyperplasia in type II cells Increases in the number of collagen bundles and edema in the alveolar septa were also noted Keywords: Deltamethrin, histopathology, Biochemical parameters, lung Number of Pages: 38 page Date of Submission: December 05th, 2016 Supervisor’s signature iii ACKNOWLEDGEMENT After an intensive period of three months; writing this note of thanks is the final statement of my thesis It has been a period of intense learning for me, not only in the scientific arena, but also on a personal level Writing this thesis has had a big impact on me I would like to reflect on the people who have supported and helped me so much throughout this period First of all, I would like to express my deepest gratitude to my supervisor, Dr.phil Arinafril of Sriwijaya University, Indralaya, Indonesia I have been amazingly fortunate to have an advisor who gave me the freedom to explore on my own, and at the same time the guidance to recover when my steps faltered He taught me how to formulate question and express ideas His valuable support helped me overcome many critical situations to finish this research work I hope that one day I would become as good an advisor to my students as Dr Arinaril has been to me I am grateful to my other supervisor Dr Nguyen The Hung for his encouragement and practical advice I am also thankful to him for reading my reports, giving important comments on my views and helping me understand and enrich my ideas In addition, I am also thankful to him for encouraging me the use of correct grammar and consistent notation in my writings and for carefully reading and commenting on countless revisions of this manuscript Ph.D Dr Krisna Murti in Department of Anatomical Pathology insightful comments and constructive criticisms at different stages of my research were thoughtprovoking and she helped me focus on my ideas I am grateful to her for holding me to iv a high research standard and enforcing strict validations for each research result, and thus teaching me how to research Dr Mulawarman from Sriwijaya University, Indralaya, Indonesia, my advisor has been always there to listen and give substantial advice I am deeply grateful to him for the long discussions and lectures on related topics that helped me improve my knowledge in the area and helped me sort out the technical details of my thesis In addition, special thanks should be offered to the Rector of Sriwijaya University, Prof Badia Perizade, for granting my internship acceptance I would also like to acknowledge with much appreciation to the Dean of Faculty of Medicine in Sriwijaya University, Dr Dr Mohammad Zulkarnain, who gave the permission to use all required equipment and the necessary materials to conduct my research in Laboratory of Department of Anatomical Pathology, Dr Mohammad Hoesin Public Hospital Particularly, I would like to acknowledge to the members of the Department of Anatomical Pathology, University of Sriwijaya/Dr Mohammad Hoesin General Hospital Palembang: Mrs Fitri Faurianty, Madi Santoso, Eka Susanti, etc… for their guidance and constant supervision as well as for providing necessary information regarding the project and also for their support in completing my research Special thanks to the technicians who work at the Animal House, Faculty of Medicine, University of Sriwijaya Especially to Mr Parman who provided me rats for free Without them, my research could not be completed on time Special thanks to all my friends from Viet Nam and Indonesia also for sharing their experiences, time and commitment especially during finishing this internship program I am grateful because I have a lot of friends were helped and supported me v throughout the course of completing the internship program My classmate: Jimlea, Ye, Keraia, Ngoc, Dung, Phong and others friends from Indonesia: Umiya, Rotua, Metri, Hendra, Irwan Jani, Dede, Naufal, Arift, Didi, Riko, etc I greatly value their friendship and I deeply appreciate their belief in me Most importantly, none of this would have been possible without the love and patience of my family My immediate familiy, to whom this research work is dedicated to, has been a constant source of love, concern, support and strength all these years I would like to express my heart-felt gratitude to my family Thai Nguyen, December 05th, 2016 Student Nguyen Thi Van vi TABLE OF CONTENT ACKNOWLEDGEMENT iv TABLE OF CONTENT vii LIST OF TABLE ix LIST OF FIGURE x PART I INTRODUCTION 1.1 Research rationale 1.2 Research Objectives 1.3 Research question PART II LITERATURE REVIEW 2.1 Deltamethrin 2.2 Toxic effects of DELTAMETHRIN on organisms 2.3 Histopathological Effects 11 2.4 Test species 12 PART III MATERIALS AND METHODS 13 3.1 Time and Place 13 3.2 Materials 13 3.3 Equipment 13 3.4 Methods 14 3.4.1 For the tissue alteration research 15 3.4.2 For histopathological examination 17 PART IV.RESULTS 18 4.1 Week 18 vii 4.2 Week 20 4.3 Week 24 PART V DISCUSSION AND CONCLUSION 27 5.1 Discussion 27 5.2 Conclusion 29 REFERENCES 31 viii LIST OF TABLE Table 1.Physical and Chemical Properties of Deltamethrin Table Toxicity classification of Deltamethrin Table Equipment used in laboratory 14 ix LIST OF FIGURE Figure 1: Bronchiole structure of control and treated rats of week 19 Figure 2: Comparison of bronchioles structure of control and treated rats 19 Figure 3: Alveoli structures of control and treated rats 20 Figure 4: The thickness of alveolar wall on the control and treated rats 21 Figure 5: Development of Bronchiolitis in treated rat 22 Figure 6: Lymp node of lung of control and treated rats 23 Figure 7: Inflammatory cells in control and treated lung structure 24 Figure 8: Swollen alveoli and lymphocyte infiltration in lung of treated rat 25 Figure 9: Structure of intrapulmonary vascular of control and treated rats 26 x a b (H&E, 10X) Figure 7: Inflammatory cells in control and treated lung structure Observed through a microscope showed (a) Normal lungs structure (b) multifocal inflammatory appear with the big size (arrow) and thickness in many places at treated lung 4.3 Week Compared to the control group, after 21 days, the lung of treated rats showed severe changes Alveolar swollen found in almost full areas of lung, more lymphocyte infiltration in the whole sites More alveolar found with increased thickness wall (Figure a, b and c) Furthermore, intrapulmonary vasculars showed much more red blood cells inside lumen with dense inflammatory cells adjacent to the arterial walls (Figure 9b) 24 a b c Figure 8: Swollen alveoli and lymphocyte infiltration in lung of treated rat (a), (b) The lung of treated group at week showed much more swollen alveoli in almost entire lung and smooth muscle hypertrophy, edema (marked by black arrow) and foci of inflammation (yellow arrow) (H&E,10X) (c) Foci of necrosis on alveolar walls causing slight disruption (black circles) (H&E,20X) 25 a b (H&E, 10X) Figure 9: Structure of intrapulmonary vascular of control and treated rats (a) Structure of intrapulmonary artery (b) The vascular of lung in treated rat showed more red blood cells (black arrow) with dense accumulation of inflammatory cells appear in vascular walls and adjacent structure (white arrow) 26 PART V: DISCUSSION AND CONCLUSION 5.1 Discussion The histological changes in rat is a noteworthy and promising field to understand the extent to which changes in the structural organization are occurring in the organs due to environmental pollution Deltamethrin exposure did significantly alter tissue structure in lung of Sprague Dawley at the concentrations tested No cells in the body operate without these tiny oxygen molecules which gives the lungs If the body is exposed to the air, there is a lot of pathogens in the environment that should penetrate the lung with many functions against the risk of hurting oneself The capillaries in the alveolar form a dense network, gas exchange duty - the main function of the lungs The capillaries go with the blood vessels that the nerves controlling the bronchial smooth muscle, dilate bronchial make or shrink The entire surface of the bronchial and alveolar mucosa with villi covered with very fine layers, always shake to get foreign objects out People often think like that the only functional lung metabolism of oxygen and CO2 In fact, every cell lung acts as a tiny plants and undertake critical functions, helps the body to sustain life Epithelial cells (covering the whole heart alveoli, bronchi) and endothelial cells (background overlaid circuit) as a barrier to water and protein molecules go too much into the interstitial tissue (organizational links between alveolar and capillary membrane), involved in the metabolism and synthesis of many important qualities In the interstitial tissue with immune cells; the number of pathological was increased when to make antibodies to help fight bacteria and strengthen its shield of the body 27 Because lung is made up by lobes, lobes separate division so when lobes are infected, the left lobe will increase capacity, making up to hurt the organization has been hurt When the body is weakened, powerful pathogens, lesions can spread to a lung or both lungs, causing severe illness During the first week (after days) After rats were cultured in the laboratory conditions for 10 days, they were put into conducted experiments From the results observed in the mouse lung tissue samples were taken after the first week of exposure by inhalation deltamethrin compared to mice kept in laboratory conditions Here are some of the observed changes in mouse lung tissue structure was exposed to drugs: an increase in inflammatory cells located around broncioles, the edema in some alveoli Second week (14 days) During this week, we can see the development of inflammatory cells, changes in fiber and have a more pronounced bleeding in the majority of bulkhead Despite the outward expression of the mouse, but nothing unusual in lung structure, but changes appeared clear Last week (21 days) Mouse lung tissue samples to look after a period of 21 days, the lungs of the mice showed changes increasingly serious: Lung cystic swelling, inflammatory cell infiltration of lymphocytes, red blood cells more inner lumen with inflammatory cells thick, etc In this study, the changes observed in the histopathology of the mouse lungs clearly shows the risks of using pesticides Although non-fatal, but during the experiment, 21 28 days continuous inhalation to insecticide (deltamethrin) for hour per day has led to very serious changes in mouse lung structures We can see the level of deltamethrin toxic if using regularly and continuously will bring unpredictable consequences, endangering the life of creatures, even affect human health in process used, inhalation is unavoidable So building the strict regulations for the indiscriminate use of pesticides is essential For a major public health and a safe application of pesticides, it is important to regularly monitor pesticide residues in food and people to assess the population exposed to pesticides as well as more testing should be done to determine the concentration and duration of exposure without causing lethal side effects significantly 5.2 Conclusion Deltamethrin is a synthetic pyrethroid with has a large-scale use, and a large number of humans and other animals are being exposed to it With this study we focused on pulmonary effects of deltamethrin in rats lung through inhalation which has not been available in the literature Findings indicate that deltamethrin can cause a variety of pulmonary disorders; therefore the use of this insecticide should be controlled seriously Deltamethrin caused degenerative changes in the lung of Sprague Dawley rat In deltamethrin poisoning greater intensity of changes was observed in male animals by microscope, with similar dose but different time to contact with deltamethrin by inhalation of rat in each group after days, 14 days and 21 days Using 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pyrethroid insecticides to Hyalella azteca: Application to field sediment with unexpectedly low toxicity Environmental Toxicology and Chemistry, 27, pp 2124-2130 35 Pictures related to this research Rat was used in this experiment The box for experiment Deltamethrin used in this research (mixed with water) The rats were taken out 36 The portions of lung rat were excised out and put on formaldehyde solution Shandon histocentre Histokinet The stained slides for lung of rat 37 The members of the Department of Anatomical Pathology, University of Sriwijaya/Dr Mohammad Hoesin General Hospital Palembang 38 ... Title : HISTOPATHOLOGICAL ALTERATIONS ON RAT AFTER EXPOSED TO DELTAMETHRIN BY INHALATION TREATEMENTS Supervisor (s): Dr Nguyen The Hung Dr Arinafril Dr Krisna Dr.Mulawarman Abstract: Deltamethrin. .. Objectives To assess the historical effects of deltamethrin on lung of rats by inhalation 1.3 Research question and hypotheses 1.3.1 Research question How does Deltamethrin affect the lung rat? 1.3.2... exposure by inhalation deltamethrin compared to mice kept in laboratory conditions Here are some of the observed changes in mouse lung tissue structure was exposed to drugs: an increase in inflammatory